Inducible Depletion of Calpain-2 Mitigates Abdominal Aortic Aneurysm in Mice

Latha Muniappan; Michihiro Okuyama; Aida Javidan; Devi Thiagarajan; Weihua Jiang; Jessica J. Moorleghen; Lihua Yang; Anju Balakrishnan; Deborah A. Howatt; Haruhito A. Uchida; Takaomi C. Saido; Venkateswaran Subramanian

doi:10.1161/ATVBAHA.120.315546

Inducible Depletion of Calpain-2 Mitigates Abdominal Aortic Aneurysm in Mice

Latha Muniappan, Michihiro Okuyama, Aida Javidan, Devi Thiagarajan, Weihua Jiang, Jessica J. Moorleghen, Lihua Yang, Anju Balakrishnan, Deborah A. Howatt, Haruhito A. Uchida, Takaomi C. Saido, Venkateswaran Subramanian

Research output: Contribution to journal › Article › peer-review

6 Scopus citations

Abstract

Objective: Cytoskeletal structural proteins maintain cell structural integrity by bridging extracellular matrix with contractile filaments. During abdominal aortic aneurysm (AAA) development, (1) aortic medial degeneration is associated with loss of smooth muscle cell integrity and (2) fibrogenic mesenchymal cells mediate extracellular matrix remodeling. Calpains cleave cytoskeletal proteins that maintain cell structural integrity. Pharmacological inhibition of calpains exert beneficial effects on Ang II (angiotensin II)-induced AAAs in LDLR-/- (low-density receptor deficient) mice. Here, we evaluated the functional contribution of fibrogenic mesenchymal cells-derived calpain-2 on (1) cytoskeletal structural protein and extracellular matrix alterations and (2) AAA progression. Approach and Results: Calpain-2 protein and cytoskeletal protein (filamin and talin) fragmentation are significantly elevated in human and Ang II-induced AAAs in mice. To examine the relative contribution of calpain-2 in AAA development, calpain-2 floxed mice in an LDLR-/- background were bred to mice with a tamoxifen-inducible form of Cre under control of either the ubiquitous promoter, chicken β-actin, or fibrogenic mesenchymal cell-specific promoter, Col1α2 (collagen type 1 alpha 2). Ubiquitous or fibrogenic mesenchymal cell-specific depletion of calpain-2 in mice suppressed Ang II-induced AAAs, filamin/talin fragmentation, while promoting extracellular matrix protein, collagen in the aortas. Calpain-2 silencing in aortic smooth muscle cells or fibroblasts reduced Ang II-induced filamin fragmentation. In addition, silencing of filamin in aortic SMCs significantly reduced collagen protein. Furthermore, calpain-2 deficiency suppressed rupture of established Ang II-induced AAAs in mice. Conclusions: Our studies implicate that calpain-2 deficiency prevents (1) Ang II-induced cytoskeletal structural protein fragmentation and AAA development and (2) stabilize and suppress rupture of established AAAs in mice.

Original language	English
Pages (from-to)	1694-1709
Number of pages	16
Journal	Arteriosclerosis, Thrombosis, and Vascular Biology
Volume	41
Issue number	5
DOIs	https://doi.org/10.1161/ATVBAHA.120.315546
State	Published - May 5 2021

Bibliographical note

Funding Information:
This study was supported by a Scientist Development Grant (14SDG18740000) from the American Heart Association and by the National Institutes of Health (Grants P20GM103527, R01HL130086). The content in this article is solely the responsibility of the authors and does not necessarily represent official views of the National Institutes of Health.

Publisher Copyright:
© 2021 American Heart Association, Inc.

Keywords

angiotensin II
calpain
cytoskeletal protein
filamin A
tamoxifen

ASJC Scopus subject areas

Cardiology and Cardiovascular Medicine

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10.1161/ATVBAHA.120.315546

Cite this

Muniappan, L., Okuyama, M., Javidan, A., Thiagarajan, D., Jiang, W., Moorleghen, J. J., Yang, L., Balakrishnan, A., Howatt, D. A., Uchida, H. A., Saido, T. C., & Subramanian, V. (2021). Inducible Depletion of Calpain-2 Mitigates Abdominal Aortic Aneurysm in Mice. Arteriosclerosis, Thrombosis, and Vascular Biology, 41(5), 1694-1709. https://doi.org/10.1161/ATVBAHA.120.315546

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title = "Inducible Depletion of Calpain-2 Mitigates Abdominal Aortic Aneurysm in Mice",

abstract = "Objective: Cytoskeletal structural proteins maintain cell structural integrity by bridging extracellular matrix with contractile filaments. During abdominal aortic aneurysm (AAA) development, (1) aortic medial degeneration is associated with loss of smooth muscle cell integrity and (2) fibrogenic mesenchymal cells mediate extracellular matrix remodeling. Calpains cleave cytoskeletal proteins that maintain cell structural integrity. Pharmacological inhibition of calpains exert beneficial effects on Ang II (angiotensin II)-induced AAAs in LDLR-/- (low-density receptor deficient) mice. Here, we evaluated the functional contribution of fibrogenic mesenchymal cells-derived calpain-2 on (1) cytoskeletal structural protein and extracellular matrix alterations and (2) AAA progression. Approach and Results: Calpain-2 protein and cytoskeletal protein (filamin and talin) fragmentation are significantly elevated in human and Ang II-induced AAAs in mice. To examine the relative contribution of calpain-2 in AAA development, calpain-2 floxed mice in an LDLR-/- background were bred to mice with a tamoxifen-inducible form of Cre under control of either the ubiquitous promoter, chicken β-actin, or fibrogenic mesenchymal cell-specific promoter, Col1α2 (collagen type 1 alpha 2). Ubiquitous or fibrogenic mesenchymal cell-specific depletion of calpain-2 in mice suppressed Ang II-induced AAAs, filamin/talin fragmentation, while promoting extracellular matrix protein, collagen in the aortas. Calpain-2 silencing in aortic smooth muscle cells or fibroblasts reduced Ang II-induced filamin fragmentation. In addition, silencing of filamin in aortic SMCs significantly reduced collagen protein. Furthermore, calpain-2 deficiency suppressed rupture of established Ang II-induced AAAs in mice. Conclusions: Our studies implicate that calpain-2 deficiency prevents (1) Ang II-induced cytoskeletal structural protein fragmentation and AAA development and (2) stabilize and suppress rupture of established AAAs in mice.",

keywords = "angiotensin II, calpain, cytoskeletal protein, filamin A, tamoxifen",

author = "Latha Muniappan and Michihiro Okuyama and Aida Javidan and Devi Thiagarajan and Weihua Jiang and Moorleghen, {Jessica J.} and Lihua Yang and Anju Balakrishnan and Howatt, {Deborah A.} and Uchida, {Haruhito A.} and Saido, {Takaomi C.} and Venkateswaran Subramanian",

note = "Funding Information: This study was supported by a Scientist Development Grant (14SDG18740000) from the American Heart Association and by the National Institutes of Health (Grants P20GM103527, R01HL130086). The content in this article is solely the responsibility of the authors and does not necessarily represent official views of the National Institutes of Health. Publisher Copyright: {\textcopyright} 2021 American Heart Association, Inc.",

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doi = "10.1161/ATVBAHA.120.315546",

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T1 - Inducible Depletion of Calpain-2 Mitigates Abdominal Aortic Aneurysm in Mice

AU - Muniappan, Latha

AU - Okuyama, Michihiro

AU - Javidan, Aida

AU - Thiagarajan, Devi

AU - Jiang, Weihua

AU - Moorleghen, Jessica J.

AU - Yang, Lihua

AU - Balakrishnan, Anju

AU - Howatt, Deborah A.

AU - Uchida, Haruhito A.

AU - Saido, Takaomi C.

AU - Subramanian, Venkateswaran

N1 - Funding Information: This study was supported by a Scientist Development Grant (14SDG18740000) from the American Heart Association and by the National Institutes of Health (Grants P20GM103527, R01HL130086). The content in this article is solely the responsibility of the authors and does not necessarily represent official views of the National Institutes of Health. Publisher Copyright: © 2021 American Heart Association, Inc.

PY - 2021/5/5

Y1 - 2021/5/5

N2 - Objective: Cytoskeletal structural proteins maintain cell structural integrity by bridging extracellular matrix with contractile filaments. During abdominal aortic aneurysm (AAA) development, (1) aortic medial degeneration is associated with loss of smooth muscle cell integrity and (2) fibrogenic mesenchymal cells mediate extracellular matrix remodeling. Calpains cleave cytoskeletal proteins that maintain cell structural integrity. Pharmacological inhibition of calpains exert beneficial effects on Ang II (angiotensin II)-induced AAAs in LDLR-/- (low-density receptor deficient) mice. Here, we evaluated the functional contribution of fibrogenic mesenchymal cells-derived calpain-2 on (1) cytoskeletal structural protein and extracellular matrix alterations and (2) AAA progression. Approach and Results: Calpain-2 protein and cytoskeletal protein (filamin and talin) fragmentation are significantly elevated in human and Ang II-induced AAAs in mice. To examine the relative contribution of calpain-2 in AAA development, calpain-2 floxed mice in an LDLR-/- background were bred to mice with a tamoxifen-inducible form of Cre under control of either the ubiquitous promoter, chicken β-actin, or fibrogenic mesenchymal cell-specific promoter, Col1α2 (collagen type 1 alpha 2). Ubiquitous or fibrogenic mesenchymal cell-specific depletion of calpain-2 in mice suppressed Ang II-induced AAAs, filamin/talin fragmentation, while promoting extracellular matrix protein, collagen in the aortas. Calpain-2 silencing in aortic smooth muscle cells or fibroblasts reduced Ang II-induced filamin fragmentation. In addition, silencing of filamin in aortic SMCs significantly reduced collagen protein. Furthermore, calpain-2 deficiency suppressed rupture of established Ang II-induced AAAs in mice. Conclusions: Our studies implicate that calpain-2 deficiency prevents (1) Ang II-induced cytoskeletal structural protein fragmentation and AAA development and (2) stabilize and suppress rupture of established AAAs in mice.

AB - Objective: Cytoskeletal structural proteins maintain cell structural integrity by bridging extracellular matrix with contractile filaments. During abdominal aortic aneurysm (AAA) development, (1) aortic medial degeneration is associated with loss of smooth muscle cell integrity and (2) fibrogenic mesenchymal cells mediate extracellular matrix remodeling. Calpains cleave cytoskeletal proteins that maintain cell structural integrity. Pharmacological inhibition of calpains exert beneficial effects on Ang II (angiotensin II)-induced AAAs in LDLR-/- (low-density receptor deficient) mice. Here, we evaluated the functional contribution of fibrogenic mesenchymal cells-derived calpain-2 on (1) cytoskeletal structural protein and extracellular matrix alterations and (2) AAA progression. Approach and Results: Calpain-2 protein and cytoskeletal protein (filamin and talin) fragmentation are significantly elevated in human and Ang II-induced AAAs in mice. To examine the relative contribution of calpain-2 in AAA development, calpain-2 floxed mice in an LDLR-/- background were bred to mice with a tamoxifen-inducible form of Cre under control of either the ubiquitous promoter, chicken β-actin, or fibrogenic mesenchymal cell-specific promoter, Col1α2 (collagen type 1 alpha 2). Ubiquitous or fibrogenic mesenchymal cell-specific depletion of calpain-2 in mice suppressed Ang II-induced AAAs, filamin/talin fragmentation, while promoting extracellular matrix protein, collagen in the aortas. Calpain-2 silencing in aortic smooth muscle cells or fibroblasts reduced Ang II-induced filamin fragmentation. In addition, silencing of filamin in aortic SMCs significantly reduced collagen protein. Furthermore, calpain-2 deficiency suppressed rupture of established Ang II-induced AAAs in mice. Conclusions: Our studies implicate that calpain-2 deficiency prevents (1) Ang II-induced cytoskeletal structural protein fragmentation and AAA development and (2) stabilize and suppress rupture of established AAAs in mice.

KW - angiotensin II

KW - calpain

KW - cytoskeletal protein

KW - filamin A

KW - tamoxifen

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Inducible Depletion of Calpain-2 Mitigates Abdominal Aortic Aneurysm in Mice

Abstract

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Keywords

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